Russell Athena L, Lefavor Rebecca C, Zubair Abba C
Transfusion Medicine, Department of Laboratory Medicine and Pathology and Center for Regenerative Medicine, Mayo Clinic, Jacksonville, Florida.
Transfusion. 2018 Oct;58(10):2374-2382. doi: 10.1111/trf.14805. Epub 2018 Sep 10.
Expanding quantities of mesenchymal stem cells (MSCs) sufficient to treat large numbers of patients in cellular therapy and regenerative medicine clinical trials is an ongoing challenge for cell manufacturing facilities.
We evaluated options for scaling up large quantities of bone marrow-derived MSCs (BM-MSCs) using methods that can be performed in compliance with Good Manufacturing Practices (GMP). We expanded BM-MSCs from fresh marrow aspirate in αMEM supplemented with 5% human platelet lysate using both an automated cell expansion system (Quantum, Terumo BCT) and a manual flask-based method using multilayer flasks. We compared MSCs expanded using both methods and assessed their differentiation to adipogenic and osteogenic tissue, capacity to suppress T-cell proliferation, cytokines, and growth factor secretion profile and cost-effectiveness of manufacturing enough BM-MSCs to administer a single dose of 100 × 10 cells per subject in a clinical trial of 100 subjects.
We have established that large quantities of clinical-grade BM-MSCs manufactured with an automated hollow-fiber bioreactor were phenotypically (CD73, CD90, CD105) and functionally (adipogenic and osteogenic differentiation and cytokine and growth factor secretion) similar to manually expanded BM-MSCs. In addition, MSC manufacturing costs significantly less and required less time and effort when using the Quantum automated cell expansion system over the manual multilayer flasks method.
MSCs manufactured by an automated bioreactor are physically and functionally equivalent to the MSCs manufactured by the manual flask method and have met the standards required for clinical application.
在细胞治疗和再生医学临床试验中，扩大间充质干细胞（MSC）的数量以满足大量患者的治疗需求，是细胞制造设施面临的一项持续挑战。
我们评估了使用符合药品生产质量管理规范（GMP）的方法扩大大量骨髓来源的间充质干细胞（BM-MSC）的选项。我们使用自动细胞扩增系统（Quantum，泰尔茂BCT）和基于多层培养瓶的手动培养方法，在添加5%人血小板裂解液的αMEM培养基中，从新鲜骨髓抽吸物中扩增BM-MSC。我们比较了使用这两种方法扩增的MSC，并评估了它们向脂肪和成骨组织的分化能力、抑制T细胞增殖的能力、细胞因子和生长因子分泌谱，以及在一项针对100名受试者的临床试验中，生产足够数量的BM-MSC以给每位受试者单次注射100×10⁶个细胞的成本效益。
我们已经确定，使用自动中空纤维生物反应器生产的大量临床级BM-MSC在表型（CD73、CD90、CD105）和功能（脂肪和成骨分化以及细胞因子和生长因子分泌）上与手动扩增的BM-MSC相似。此外，与手动多层培养瓶方法相比，使用Quantum自动细胞扩增系统时，MSC的制造成本显著降低，所需时间和精力更少。
通过自动生物反应器制造的MSC在物理和功能上等同于通过手动培养瓶方法制造的MSC，并且已达到临床应用所需的标准。
